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| 0.99 | The title is: 迷迭香中樟脑的起源及其生物合成机制研究 |
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Title: 迷迭香中樟脑的起源及其生物合成机制研究
迷迭香中樟脑的组织分布与含量变异特征
迷迭香樟腦在不同組織與器官中的分布特徵
迷迭香樟腦的組織與器官分布具有明顯差異，且含量存在一定變異特徵，其差異主要體現在組織器官類型、季節、生態型及樣品來源等多個維度。
迷迭香樟腦含量的組織器官與來源差異彙總
| 分類維度 | 具體對象 | 樟腦含量/比例信息 | 文獻來源 |
|---|---|---|---|
| 器官比例 | 果實 | 樟腦與1,8-桉葉素比例為4:1 | [1] |
| 器官比例 | 老枝條 | 樟腦與1,8-桉葉素比例為2:1 | [1] |
| 器官比例 | 嫩枝條 | 樟腦與1,8-桉葉素比例接近1:1 | [1] |
| 葉片揮發油 | 綠葉 | 主要萜類成分，含量超過90% | [2] |
| 葉片揮發油 | 落葉 | 主要萜類成分，含量約80-90% | [2] |
| 葉片精油 | 栽培樣品 | 4.3% | [3] |
| 葉片精油 | 部分樣品 | 10.5% | [4] |
| 葉片精油 | 部分樣品 | 12.41% | [5] |
| 葉片精油 | 部分樣品 | 26.30% | [6] |
| 花朵提取物 | 正己烷萃取物 | 檢測到樟腦存在 | [7] |
| 花朵揮發物 | 揮發性物質 | 含有樟腦 | [8] |
| 季節性變化（嫩枝） | 5月採集 | 18.8%（含量最低） | [1] |
| 季節性變化（老枝） | 全年 | 23.3%-28.1%（含量穩定） | [1] |
| 環境樣品 | 土壤 | 主要萜類成分之一 | [2] |
| 環境樣品 | 水溶膠頂空成分 | 5.4% | [9] |
| 精油來源 | 部分精油 | 9.38% | [10] |
| 精油來源 | 精油 | 11.7% | [11] |
| 精油來源 | 水劑 | 15.3% | [11] |
| 精油來源 | 部分精油 | 14.12% | [12] |
| 精油來源 | 部分精油 | 15.2% | [13] |
| 精油來源 | 部分精油 | 19% | [14] |
| 精油來源 | 部分精油 | 20.42% | [15] |
| 精油來源 | 部分精油 | 26.30% | [6] |
| 生態型 | Cevoli生態型精油平均 | 9.1% | [16] |
| 生態型 | Lunigiana生態型精油平均 | 5.7% | [16] |
| 植株部位 | 中間部位 | 8.2%（最高） | [16] |
| 植株部位 | 下部 | 7.4% | [16] |
| 植株部位 | 頂部 | 6.7% | [16] |
| Cevoli生態型器官 | 下部、中間部位花朵 | 含量最高 | [16] |
| Cevoli生態型器官 | 各部位莖 | 含量最低 | [16] |
| Lunigiana生態型器官 | 中間、下部莖、葉 | 含量最豐富 | [16] |
| Lunigiana生態型器官 | 頂部花朵 | 2.5%（最低） | [16] |
| 精油主要成分 | 部分精油 | 52%（w/w，主要成分） | [17] |
| 精油含量多樣性 | 部分精油 | 15.57% | [18] |
| 精油含量多樣性 | 部分精油 | 23.24% | [19] |
| 精油含量多樣性 | 部分精油 | 22.58% | [20] |
| 精油含量多樣性 | 部分精油 | 24.66% | [21] |
| 精油含量多樣性 | 部分精油 | 26.31% | [22] |
| 精油含量多樣性 | 部分精油 | 最高可達18% | [23] |
樟腦含量的變異特徵不僅體現在組織器官層面，還受到季節、生態型及植株部位等因素的影響。例如，嫩枝條樟腦含量呈現季節性波動，5月採集樣品含量最低；老枝條則全年維持穩定水平[1]。不同生態型間的差異明顯，Cevoli生態型精油平均樟腦含量高於Lunigiana生態型，且兩者在植株不同部位的含量分布模式也存在區別[16]。此外，迷迭香精油中樟腦含量呈現高度多樣性，部分研究顯示其可作為主要成分（含量達52% w/w），而不同來源、生態型的精油含量範圍覆蓋5.7%至26.31%不等。值得注意的是，樟腦不僅存在於迷迭香植株組織中，還可在土壤、水溶膠等環境樣品中檢測到，進一步體現其分布的廣泛性[27,96]。迷迭香揮發油的化學特徵由樟腦與α-蒎烯、1,8-桉葉素、龍腦等多種成分共同構成[106,178,188]。
迷迭香樟腦含量的品種與地理變異規律
迷迭香精油中樟脑的含量存在显著的品种、地理、生态型、季节、组织部位及提取方法变异特征，且与化学型分类密切相关。不同来源样品的樟脑含量差异较大，部分研究明确了品种与地理因素的影响：意大利品种（如Gorizia品种13.02%）显著高于法国品种（2.69%）[24]；伊朗不同种群樟脑含量范围为1.66%至24.82%[25]，另有报道为7.21%[26]；摩洛哥和突尼斯属于樟脑占优势的化学型[27]，西班牙和突尼斯含量分别为9.2%和9.7%[28]；摩洛哥不同地区（Middle Atlas和Loukkos）因含氧单萜类占比差异，樟脑含量也随地区变化[29]。生态型间差异同样明显，如Cevoli生态型（9.1%）高于Lunigiana生态型（5.7%）[16]，且两者组织部位分布特征不同：Cevoli生态型下部和中部花含量最高、茎最低[16]；Lunigiana生态型中部和下部茎与叶最高、顶部花最低（2.5%）[16]，整体中部组织含量最高（8.2%）[16]。
季节与收获因素影响显著：2月（开花前）樟脑浓度高于其他季节[30]；秋季含量最高，且品种与收获季节存在交互作用[31]。化学型分类中，部分精油以樟脑为主（如Cluster 3达35.6%、var. '8'为“樟脑型”[32]），或与1,8-桉叶素、α-蒎烯含量相近（20-30%）[33]。品种/品系及克隆系差异也较突出：IHBT/RMAc-1含量显著高于IHBT/RMAc-2[31]；Pigette和Esselte克隆樟脑为主要成分（前者达25%），Nonza克隆仅5%[34]。提取方法中，水蒸气蒸馏法平均含量高于溶剂萃取法[32]。这些变异可能与环境调控植物代谢[30]、遗传因素、化学型及营养状况有关[35]。
迷迭香精油樟脑含量的样品来源差异汇总
| 样品来源特征 | 樟脑含量 | 参考文献 |
|---|---|---|
| 高含量样品 | 25.22%、20.42%、22.62%、13.2%、13.09%、27.7% | [36][15][33][37][38][39] |
| 高含量样品 | 41.22% | [40] |
| 低含量样品 | 0.7-5.8%、9.3%、9.3%、5%、9.2%、9.7% | [30][41][42][43][28] |
| 意大利Gorizia品种 | 13.02% | [24] |
| 法国品种 | 2.69% | [24] |
| 伊朗种群 | 1.66%-24.82% | [25] |
| 伊朗精油 | 7.21% | [26] |
| 摩洛哥化学型样品 | 40.0 mg/mL | [27] |
| 样品1-12 | 32.2-41.2% | [44] |
| 样品13 | 3.1% | [44] |
| 样品14-15 | 11.1-13.4% | [44] |
| 迷迭香叶提取精油 | 23.04% | [45] |
| 部分样品范围 | 10.7%-32.9% | [46] |
| 不同地区样品 | 12.07%、10.43%、16.74%、17.56%、18.9%、14.80%、24.66% | [47][48][49][50][51][52][53] |
| 部分样品 | 22.21% | [54] |
迷迭香樟腦含量的季節與採收期變異特點
迷迭香樟腦的組織分布與含量受多種因素影響，包括組織類型、種質資源、採收時期及年份等，整體呈現明顯差異。樟腦是迷迭香不同器官精油中的主要成分，但其與1,8-桉葉素的含量比值及具體含量範圍均因組織類型而異；同時，不同種質資源、季節與年份的樟腦含量也存在顯著變異。
迷迭香不同組織類型的樟腦含量及與1,8-桉葉素的比值
| 組織類型 | 樟腦與1,8-桉葉素的比值 | 樟腦含量範圍 | 文獻來源 |
|---|---|---|---|
| 果實 | 約4:1 | - | [1] |
| 老枝條 | 2:1 | 23.3-28.1% | [1] |
| 嫩枝條 | 接近1:1 | 18.2-18.8% | [1] |
| 新鮮地上部分全株 | - | 23.9-33.2% | [55] |
| 葉片 | - | 24.8-35.8% | [55] |
迷迭香樟腦含量的其他影響因素
| 影響因素 | 具體內容 | 文獻來源 |
|---|---|---|
| 種質資源 | IHBT/RMAc-1品系樟腦含量顯著高於IHBT/RMAc-2品系 | [31] |
| 種植合作社 | C1合作社（7.3±0.5%）顯著高於C2合作社（5.8±0.2%）和C3合作社（5.3±0.1%） | [56] |
| 季節趨勢（整體） | 冬季（Group A）含量較高，夏季（Group B）顯著降低 | [56] |
| 季節趨勢（嫩枝條） | 5月（採收期）最低，為18.8% | [1] |
| 季節趨勢（葉片） | T1時期最高（35.8%），T5時期最低（24.8%） | [55] |
| 季節趨勢（地上部分） | T1時期最高（33.2%），T6時期最低（23.9%） | [55] |
| 季節與品系交互作用 | 秋季採收的IHBT/RMAc-2品系含量最高，其次是同季節的IHBT/RMAc-1品系 | [31] |
| 年份差異 | 2011年為6.2%，2012年為15.3% | [57] |
上述結果表明，迷迭香樟腦含量的變異是組織特性、遺傳背景與環境因子共同作用的結果，這些差異為迷迭香資源的定向開發（如高樟腦品系選育、最佳採收時期確定）提供了重要依據。
迷迭香樟腦在腺毛中的定位與積累動態
迷迭香叶片中樟脑的组织定位、含量特征及环境与生理相关性可通过多种技术手段揭示。组织化学染色与色谱分析的结合明确了樟脑的合成位点与含量占比，而环境因子与生理过程的研究则进一步阐明了其积累与排放规律。
迷迭香叶片中樟脑的合成位点、含量及相关影响因素
| 研究内容 | 具体结果 | 参考文献 |
|---|---|---|
| 组织定位（染色技术） | 2,4-二硝基苯肼染色显示盾状腺毛呈轻微阳性反应（橙棕色），含羰基萜类主要由其产生 | [58] |
| GC分析（里约热内卢样本） | 樟脑为主要含羰基萜类，占挥发性化合物总量23.2%，是含量最高的单萜类成分 | [58] |
| 竞争环境影响 | 与地中海松共生时，樟脑积累量呈降低趋势（ANOVA，p = 0.06） | [59] |
| 排放与浓度相关性 | 排放因子与特化结构中樟脑浓度线性相关（p < 0.05，r = 0.50） | [59] |
| 樟脑排放比例 | 叶片中平均每小时约1.10%的樟脑储备通过排放进入大气 | [59] |
上述结果表明，盾状腺毛是迷迭香叶片樟脑合成的关键结构，其含量不仅受共生植物的竞争影响，还通过特化结构中的浓度调控向大气的排放过程，为深入理解迷迭香樟脑的代谢调控机制提供了多维度证据。
迷迭香樟腦提取與分析方法的參數優化
迷迭香精油中的樟脑含量受种质资源、提取方法及处理条件的显著影响，常与1,8-桉叶素、α-蒎烯等共同构成精油主要组分。不同因素对樟脑含量的具体影响及对应研究结果如下表所示：
迷迭香樟脑含量的影响因素及相关研究结果
| 影响因素 | 具体条件/类型 | 樟脑含量/结果 | 文献来源 |
|---|---|---|---|
| 分析方法 | HS-SPME/GC-MS | 25.22% | [36] |
| 分析方法 | 未明确（另一研究） | 24.31% | [60] |
| 种质资源 | Cluster 3中的7个基因型 | 高达35.6% | [32] |
| 干燥方式 | 冷冻干燥（顶空成分） | 9.8% | [61] |
| 干燥方式 | 喷雾干燥 | 含氧化合物（含樟脑）丰度升高 | [61] |
| 居群差异 | 居群1（Hys） | 20.3% | [62] |
| 居群差异 | 居群2（Hys） | 18.4% | [62] |
| 提取方法 | 水蒸气蒸馏法 | 平均含量高于溶剂萃取法 | [32] |
| 提取参数（MAI） | 400 W功率，处理10 min | 得率达12.39%（显著高于传统浸提法12 h的得率，后者不足前者一半） | [36] |
| 提取参数（蒸馏） | 新鲜样品，蒸馏160 min | 浓度最高（15.9%）；高沸点的樟脑在80-160 min较长蒸馏时间下浓度更高 | [63] |
此外，樟脑的分析可通过多种技术实现：GC-MS可检测其特征碎片离子（m/z 152、108、95等）[60][64]；FT-IR光谱中1744 cm⁻¹处的C=O伸缩振动峰可作为鉴定依据[64]；采用Elite-Betacydex环糊精毛细管柱的GC方法可分离单萜对映体，用于叶片中樟脑的定量分析，其含量可达9.1 mg/g干重[65]。
迷迭香樟脑含量的环境与栽培调控因素
迷迭香中樟脑的含量受多种环境与栽培因素调控，这些因素通过影响基于龙脑基二磷酸合成与转化的樟脑生物合成途径发挥作用[66]，同时其差异还与遗传背景、化学型、地理位置、采收时间、蒸馏技术等因素相关[67][35][68]，环境因素更可通过激活或抑制精油合成相关酶基因的表达调控樟脑代谢，这种调控可能涉及DNA甲基化、组蛋白修饰等表观遗传机制[69]。以下将不同因素对樟脑含量的具体影响整理如下：
不同因素对迷迭香樟脑含量的影响
| 影响因素 | 具体条件/处理 | 樟脑含量变化及特征 | 参考文献 |
|---|---|---|---|
| 季节 | 雨季采收 | 含量（25.85%）显著高于其他季节 | [70] |
| 季节 | 冬季、秋季（2月、11月峰值） | 显著高于夏季、春季，在2月和11月达到峰值 | [71] |
| 季节 | 高温月份（如1月） | 积累量最大 | [72] |
| 季节 | 秋季采收（IHBT/RMAc-2品种） | 含量最高，其次是同一季节的IHBT/RMAc-1；秋季显著高于夏季和雨季 | [31] |
| 温度 | 与月平均温度 | 呈正相关（r=0.75） | [73] |
| 温度 | 较冷条件 | 含量升高（与1,8-桉叶素响应趋势相反） | [74] |
| 光照 | 暗培养幼叶（最幼嫩叶片） | 樟脑等药理活性成分比例较低 | [75] |
| 生物胁迫 | 真菌侵染+较高剂量5-氨基乙酰丙酸（ALA） | 进一步增强真菌侵染下樟脑的积累 | [76] |
| 栽培措施 | 1.5 L ha⁻¹ Fosnutren处理 | 比氮磷钾（N.P.K.）处理高33%（p ≤ 0.01） | [77] |
| 栽培措施 | 纳米肥料处理组 | 显著高于对照组 | [78] |
| 干燥方式 | 冷冻干燥 | 比冷冻样品更能保留樟脑 | [79] |
| 干燥方式 | 干燥时间延长 | 可能因转化为乙酸龙脑酯而降低 | [80] |
| 提取方法 | 超临界二氧化碳萃取（SFE） | 含量（15.7%）高于水蒸馏（HD，9.0%） | [81] |
| 提取方法 | 甘油预处理水蒸馏 | 主要成分定性组成未变，仅定量存在差异 | [82] |
| 提取方法 | 水蒸气蒸馏（CH）vs微波萃取（ME） | 水蒸气蒸馏更利于樟脑积累（野生迷迭香WR表现优于栽培品种CR） | [83] |
| 土壤盐分 | 100 mM NaCl处理 | 显著提高相对丰度 | [84] |
| 土壤盐分 | 100 mM NaCl + 0.1 mM FeCl₃处理 | 含量降低80% | [84] |
| 品种/化学型 | 野生迷迭香（WR）vs栽培品种（CR） | WR表现优于CR | [83] |
| 品种/化学型 | 樟脑型群体vs1,8-桉叶素型群体 | 樟脑型含量显著更高 | [74] |
| 土壤类型 | 非气候性弱发育岩性土/粗骨土 | 13个群体中含量最高（范围5.88%-27.95%） | [85] |
| 土壤类型 | 石灰镁质土壤（如黑色石灰土） | 含量较低 | [85] |
| 植物生长调节剂 | 0.5 mg L⁻¹ 2,4-二氯苯氧乙酸（2,4D） | 愈伤组织中含量（8.25%）高于1.0 mg L⁻¹处理（6.05%） | [86] |
| 养分条件 | 叶片氮含量、磷含量 | 总萜类浓度与氮含量呈正相关（p < 0.001），与磷含量呈正相关（p < 0.05） | [65] |
| 水分胁迫 | 近红外（NIR）处理 | 精油中樟脑含量为14.581% | [87] |
| 水分胁迫 | 土壤水分（SW）处理 | 精油中樟脑含量为13.720% | [87] |
| γ射线辐照 | 1000 Gy辐照 | 含量从1.4%升至3.2%，相对比例显著提高 | [88] |
此外，不同地理群体的樟脑含量差异显著，如突尼斯群体为8.5%至30.17%[89]，西班牙13个野生群体为14.4%至30.7%（Alcarria地区群体较高）[90]，平原地区群体（20.86%）显著高于山区（19.21%）[91]；13个迷迭香群体的樟脑含量范围为5.88%-27.95%，其中生长在非气候性弱发育岩性土或粗骨土的群体含量最高，石灰镁质土壤（如黑色石灰土）群体含量较低[85]。化学型方面，樟脑型群体的樟脑含量显著高于1,8-桉叶素型[74]。季节与品种的交互作用对樟脑含量有显著影响，如IHBT/RMAc-2在秋季采收时樟脑含量最高，其次是同一季节的IHBT/RMAc-1，且秋季采收的迷迭香樟脑含量显著高于夏季和雨季[31]。水分胁迫下，近红外（NIR）和土壤水分（SW）处理的迷迭香精油中均检测到樟脑，其中NIR处理含量更高[87]；γ射线辐照处理会显著提高樟脑的相对比例[88]。这些因素共同构成了迷迭香樟脑含量调控的复杂网络。
迷迭香樟脑相关的化学型分类与地理分布规律
迷迭香的化学型分类通常以樟脑、α-蒎烯、1,8-桉叶素等主要挥发性成分的相对含量为依据，聚类分析是鉴定化学型的常用方法。不同研究基于主要成分的相对含量或聚类结果，划分出包含樟脑的多种化学型；同时，地理来源、提取方法及环境因素会影响樟脑含量，进而影响化学型特征。
迷迭香中含樟脑的化学型分类及相关特征
| 研究依据/背景 | 化学型类型及特征 | 参考文献 |
|---|---|---|
| 聚类分析（肘部法） | 樟脑型（C2，含60%樟脑）、α-蒎烯/樟脑/1,8-桉叶素型（C4） | [92] |
| 主要成分相对含量 | 樟脑型、樟脑/1,8-桉叶素/龙脑型、1,8-桉叶素/樟脑型 | [93] |
| 特定种质资源聚类分析 | 樟脑为主导成分（含量35.6%）的聚类群，包含多个基因型 | [32] |
| 精油主要化合物种类 | 樟脑型（以樟脑为主要成分）、1,8-桉叶素型、α-蒎烯型等 | [94][95] |
| 四类化学型划分 | 樟脑型（原产于法国、西班牙、希腊、意大利和保加利亚） | [94] |
| 混合化学型识别 | 含35–45% 1,8-桉叶素和21–23%樟脑的混合类型、樟脑/1,8-桉叶素型 | [96][97] |
| 聚类分析+PCA（塞尔维亚和黑山） | 樟脑型（Belgrade、Lusˇtica种群）、1,8-桉叶素/樟脑中间型（Zakynthos种群）；樟脑与1,8-桉叶素的温度响应呈相反趋势 | [74] |
| 西班牙安达卢西亚种群 | 樟脑含量较高，与月桂烯共同构成“化学型C”，二者呈显著正相关（r = 0.72；P ≤ 0.01） | [98] |
| 印度斯利那加地区 | 樟脑含量38.66%，为樟脑化学型 | [99] |
| 突尼斯干旱生境种群 | Matmata和Toujène地区（R. officinalis var. troglodytorum）樟脑含量25.7–30.2% | [89] |
| 不同国家品种比较 | 意大利Gorizia品种樟脑含量13.02%，显著高于法国品种的2.69%；摩洛哥品种为9.177% | [24][100] |
| 阿尔及利亚撒哈拉地区 | Béchar、Adrar等省部分样品樟脑含量达31.0% | [101] |
| 部分低樟脑含量样品 | 樟脑相对浓度不超过14.10% | [102] |
| 西班牙精油成分特征 | 樟脑和龙脑含量较高，1,8-桉叶素含量较低 | [95] |
| 摩洛哥和突尼斯精油 | 常以1,8-桉叶素为主要成分，但部分样品樟脑含量较高（如突尼斯Matmata和Toujane地区） | [97] |
| 摩洛哥樟脑含量范围 | 2.6–30.5% | [94] |
| 提取方法影响 | 水蒸气蒸馏法提取的精油中樟脑平均含量高于溶剂萃取法 | [32] |
地理来源是樟脑含量差异的重要因素，如印度斯利那加地区樟脑含量高达38.66%，而摩洛哥部分样品仅2.6%；环境因素（如温度）也会调控樟脑与其他成分的比例，例如樟脑在较冷条件下含量升高，与1,8-桉叶素的温度响应呈相反趋势。此外，提取方法对樟脑含量有直接影响，水蒸气蒸馏法提取的精油樟脑平均含量高于溶剂萃取法[32]。
迷迭香樟脑生物合成的前体途径与关键反应
迷迭香中樟脑的生物合成以香叶基焦磷酸为起始物质，需经多步反应完成：首先香叶基焦磷酸经芳樟基焦磷酸环化生成龙脑基焦磷酸，随后通过水解反应转化为龙脑，最终经龙脑羟基的氧化反应生成樟脑[103][95]。龙脑基焦磷酸的合成与转化是樟脑、龙脑及1,8-桉叶素生物合成的共同关键步骤，环境条件会影响这些复杂反应的进程，进而决定次级代谢产物的含量与质量[66]。此外，酸性迷迭香水溶胶（pH 4.2–4.5）中存在樟脑还原为龙脑的逆反应现象[104]。
迷迭香樟脑的提取方法与定量分析技术
迷迭香樟脑的种质资源与化学型分布特征
迷迭香挥发油中樟脑的存在形式与含量具有丰富的多样性，其分布特征受提取方法、处理条件、种质资源化学型及地理来源等多因素影响。在迷迭香挥发物的顶空成分中，(R)-(+)-樟脑是8种能引发触角反应的萜类化合物之一[105]。不同提取方法对樟脑含量的影响显著，且微波辅助蒸馏（MHDG）和亚临界流体萃取（SFME）等方法所得挥发油中樟脑等含氧单萜类化合物含量更高，对应更高品质的挥发油[106]。此外，不同处理条件也会改变樟脑含量，如100%水提物处理的迷迭香挥发油中樟脑含量为8.9%，而100%丙酮提取物处理的则为10.30%[107]。
迷迭香挥发油提取方法与樟脑含量的比较
| 提取方法 | 樟脑含量特征 | 参考文献 |
|---|---|---|
| 常规水蒸气蒸馏法（HD） | 含量为1.22%（干基得率1.8%） | [106] |
| 超临界CO₂萃取（SC-CO₂） | 显著低于HD法，仅为HD法的25% | [108] |
| 水蒸气蒸馏法 vs 溶剂萃取法 | 平均含量高于溶剂萃取法，且挥发性相互作用法证实差异极显著（***P < 0.0001） | [32] |
迷迭香樟脑的含量在不同样本中呈现较大波动，范围从7.94%至41.22%不等[109][40]，具体含量与种质资源的化学型分布密切相关。基于挥发油化学成分，迷迭香可分为α-蒎烯型、1,8-桉叶素型、樟脑型和马鞭草酮型四种化学型[106]；通过K-means聚类分析，其化学型可进一步细分为“樟脑型”（如var. '8'）和“1,8-桉叶素+樟脑型”（如var. '2'）等类型，其中Cluster 3以樟脑（35.6%）为主导，包含7个迷迭香种质基因型（var. '6'、'8'、'H7'、'H8'、'H14'、'H15'和'H17'）[32]。不同地理来源的迷迭香化学型特征不同，如西班牙型迷迭香挥发油中樟脑含量为13–21%，摩洛哥和突尼斯型则为5–15%[106]。此外，迷迭香挥发油可根据主要成分分为两类：一类含超过40%的1,8-桉叶素，另一类则含20–30%的1,8-桉叶素、α-蒎烯和樟脑[33]。樟脑型迷迭香挥发油的典型特征为樟脑含量占主导，如某样本中樟脑含量达41.22%，同时伴随较高的莰烯（18.14%）和α-蒎烯（17.49%）含量[40]。
不同提取方法对迷迭香樟脑产量的影响及动力学变化
迷迭香樟脑的提取方法多样，涵盖传统水蒸馏（HD）、微波辅助提取（微波辅助水扩散与重力法MHD、微波辅助浸渍MAI）、亚临界流体萃取（SFME）、超临界CO₂萃取（SC-CO₂）及索氏提取等，不同方法对樟脑产量及动力学特性影响显著[106][110][111][108][112]。各提取方法的樟脑产量差异明显，具体数据如下：
不同提取方法的迷迭香樟脑产量对比
| 提取方法 | 关键参数 | 樟脑产量/含量 | 对比结果 | 参考文献 |
|---|---|---|---|---|
| 水蒸馏（HD） | - | 干基精油得率1.8%，樟脑含量1.22% | HD法樟脑平均含量显著高于溶剂萃取法 | [106][32] |
| 微波辅助浸渍（MAI） | 400W功率，10分钟 | 12.39% | 显著高于传统浸渍12小时（未达前者一半） | [36] |
| 微波辅助浸渍（MAI） | 400W功率，4分钟 | 0.45% | 与传统浸渍12小时相当 | [36] |
| 微波辅助浸渍（MAI） | 400W功率，5分钟 | 2.9% | - | [36] |
| 微波辅助水扩散与重力法（MHD） | - | 含氧单萜类（含樟脑）含量34.3% | 高于HD法的32.2%（减少热水解效应） | [110] |
| 亚临界流体萃取（SFME） | 30分钟 | 樟脑含量12.86%，含氧单萜类含量28.6% | 樟脑含量高于HD法（5分钟峰值10.10%后下降），含氧单萜类含量高于HD法的26.98% | [111][113] |
| 超临界CO₂萃取（SC-CO₂） | - | 樟脑提取量为HD法的25% | 樟脑浓度为索氏提取的18倍 | [108][112] |
| 索氏提取 | - | 0.5±0.1g/100g提取物 | 樟脑浓度显著低于SC-CO₂萃取 | [112] |
| 微波辅助水扩散与重力法（MHG） | - | 17.51% | 与HD法（17.33%）相近 | [114] |
樟脑提取动力学因方法而异：SFME法中，含氧单萜类（含樟脑）提取量随时间逐渐增加，30分钟时达36.76%[111]；HD连续提取中，樟脑含量在5分钟达峰值10.10%后逐渐降低，而HD间歇提取中，樟脑所在的含氧单萜类含量在30分钟达峰值29.41%，3小时时降至21.78%[111]；超临界萃取中，樟脑在提取初期即被大量萃取，床层几何结构影响动力学——E-2床层中樟脑在液固比（S/F）<4时几乎耗尽，E-1床层则需S/F>4才耗尽[112]。此外，提取时间（DT）对樟脑浓度影响显著：新鲜原料在160分钟DT时樟脑浓度最高（15.9%），高沸点的樟脑在80-160分钟DT时浓度更高[63]；新鲜原料樟脑提取符合米氏方程，最大产量为63.5mg，半最大产量时间为2.11分钟[63]。
樟脑的定量分析技术主要包括气相色谱-火焰离子化检测器（GC-FID）、气相色谱-质谱联用（GC-MS）及顶空固相微萃取（HS-SPME）等[36][115][111][112][116]：GC-FID通过外标法校准曲线定量，使用DB-5毛细管柱分离并对比标准品保留时间鉴定樟脑[112]；GC-MS通过与标准品保留指数及质谱库比对鉴定樟脑[36][111]；HS-SPME/GC-MS可检测到迷迭香精油中樟脑含量为25.22%，且测得的樟脑浓度高于蒸馏法，精密度也更高（变异系数0.13-0.64，蒸馏法为1.4-5.5）[116]。
樟脑含量还受种质资源、季节及提取条件波动影响：樟脑型迷迭香樟脑含量可达35.6%，西班牙型为13-21%，摩洛哥和突尼斯型为5-15%[106][32]；某研究中樟脑含量随季节变化范围为24.38-35.93%[117]；部分研究显示不同提取条件下樟脑含量波动范围为14.47-20.4%[118]。
迷迭香樟脑的气相色谱-质谱联用定量分析技术
迷迭香樟脑的气相色谱-质谱联用（GC-MS）定量分析技术是其定量研究的核心手段之一，在迷迭香挥发油或提取物的成分鉴定与定量中应用广泛。具体分析方法方面，有研究采用GCMS-QP2010 plus系统结合DB-5ms色谱柱（30 m × 0.25 mm I.D. × 0.25 µm df），通过程序升温（初始60℃保持4 min，以3℃/min升至100℃，1℃/min升至110℃，5℃/min升至150℃，15℃/min升至300℃并保持25 min）、进样量1.6 µL、分流比1:10、进样口温度250℃、He载气（流速36.4 cm·s−1）的色谱条件，以及接口温度280℃、离子源温度230℃、质量范围m/z 40–500、扫描速度2500 amu/s、事件时间0.20 s的质谱参数进行定量分析[119]。化合物鉴定通过Wiley质谱数据库匹配及C8–C30烷烃混合物测定的线性保留指数（LRI）验证，定量则利用樟脑标准品建立2.5–25.0 µg/mL的校准曲线[119]。此外，顶空固相微萃取-气相色谱-质谱联用（HS–SPME/GC–MS）也可用于检测迷迭香挥发油中的樟脑[120]，其存在形式为(R)-(+)-樟脑[105]；迷迭香挥发油中樟脑的对映体拆分可通过GC-MS实现，能与α-蒎烯、莰烯等共分离出8对可检测的对映异构体[116]。在定量方法的线性响应方面，PDMS纤维对樟脑的线性响应良好（相关系数0.9760）[116]；采用内标法时，樟脑回归方程的R²为0.9702–0.9866[121]。部分研究中，樟脑的定量需结合标准品：通过与樟脑标准品（CAS 76-22-2）的保留指数对比鉴定，并利用外标校准曲线完成定量[112]。
不同批次或类型的迷迭香样品经GC-MS分析后，樟脑含量存在明显差异，具体数据如下：
迷迭香样品中樟脑含量的GC-MS分析结果
| 样品类型 | 樟脑含量 | 文献来源 |
|---|---|---|
| 迷迭香地上部分提取物 | 主要峰成分之一 | [122] |
| 迷迭香挥发油 | 18.7% | [123] |
| 迷迭香挥发油 | 12.41% | [5] |
| 迷迭香挥发油 | 20.42% | [15] |
| 迷迭香挥发油 | 9.3% | [41] |
| 迷迭香挥发油 | 7.94% | [109] |
| 迷迭香挥发油 | 5.11% | [124] |
| 迷迭香挥发油 | 8.97% | [125] |
| 迷迭香挥发油 | 39.46% | [126] |
| 迷迭香提取物 | 8.6–50.3 mg/g | [119] |
| 迷迭香cineole型挥发油 | 8.38% | [127] |
| 迷迭香提取物 | 0.8–6.7 mg/g | [116] |
| 某迷迭香挥发油 | 12.53% | [128] |
| 迷迭香挥发油 | 10–17.92% | [129] |
| 某迷迭香样品 | 0.308 g/L | [121] |
迷迭香樟脑在精油中的主要成分地位及地域含量差异
迷迭香精油的核心组分为樟脑及α-蒎烯、桉叶素（1,8-桉叶素）、龙脑等单萜类化合物[122][130][131][132][133][106][36][123][5][15][33][42][40][115]，其主要成分还包括莰烯、对伞花烃、月桂烯、β-蒎烯、乙酸龙脑酯等[134]。樟脑作为迷迭香精油的关键组分，其含量受来源类型、化学型、提取方法等多种因素影响，不同研究中樟脑的具体占比存在显著差异。
迷迭香精油中樟脑含量的差异（按影响因素分类）
| 影响因素类型 | 具体条件/来源 | 樟脑含量（占比/得率） | 参考文献 |
|---|---|---|---|
| 来源类型 | 西班牙类型 | 13–21% | [106] |
| 来源类型 | 摩洛哥和突尼斯类型 | 5–14% | [106] |
| 来源类型 | 撒丁岛类型 | 主要组分（未提及具体数值） | [113] |
| 来源类型 | 某伊朗迷迭香精油 | 4.97% | [117] |
| 来源类型 | 另一伊朗迷迭香精油 | 37.60% | [135] |
| 来源类型 | 比利时产樟脑型精油 | 主要含氧单萜成分（未提及具体数值） | [136] |
| 来源类型 | 某商业迷迭香精油 | 35.5% | [117] |
| 来源类型 | 迷迭香地上部分提取精油 | 主要峰成分之一（19号峰） | [122] |
| 化学型 | A型迷迭香精油 | 7.1% | [137] |
| 化学型 | B型迷迭香精油 | 未提及 | [137] |
| 提取方法 | 微波辅助浸渍（MAI，400 W，10 min） | 12.39%（橄榄油中得率） | [36] |
| 提取方法 | 传统浸渍（12 h） | 低于MAI法（未提及具体数值） | [36] |
| 提取方法 | 水蒸气蒸馏（SD） | 14.26% | [138] |
| 提取方法 | 水蒸馏 | 14.63%–39.46% | [126] |
| 提取方法 | 乙醇提取物（Et-E） | 19 mg/100 g Et-E | [139] |
| 提取方法 | 水提取物（Aq-E） | 2.4 mg/100 g Aq-E | [139] |
| 其他研究报道 | 某迷迭香精油 | 41.22% | [40] |
| 其他研究报道 | 某迷迭香精油 | 26.5–26.6% | [130] |
| 其他研究报道 | 某迷迭香精油 | 14.94% | [131] |
| 其他研究报道 | 某迷迭香精油 | 16.7% | [133] |
| 其他研究报道 | 某迷迭香精油 | 18.7% | [123] |
| 其他研究报道 | 某迷迭香精油 | 12.41% | [5] |
| 其他研究报道 | 某迷迭香精油 | 20.42% | [15] |
| 其他研究报道 | 某迷迭香精油 | 22.62% | [33] |
| 其他研究报道 | 某迷迭香精油 | 9.3–10.5% | [42] |
| 其他研究报道 | 某迷迭香精油 | 10.91% | [115] |
| 其他研究报道 | 某迷迭香精油 | 39.46% | [126] |
| 其他研究报道 | 某迷迭香精油 | 26.30% | [140] |
| 其他研究报道 | 某迷迭香精油 | 14.26% | [138] |
| 其他研究报道 | 某迷迭香精油 | 10.8% | [141] |
| 其他研究报道 | 某迷迭香精油 | 14.39%–17.17% | [117] |
| 其他研究报道 | 某迷迭香精油 | 24.31% | [60] |
| 其他研究报道 | 某迷迭香精油 | 26.31% | [22] |
| 其他研究报道 | Ro-B样品精油 | 约11% | [142] |
| 其他研究报道 | 某迷迭香精油 | 25% | [143] |
| 其他研究报道 | 某迷迭香精油 | 17.92% | [129] |
| 其他研究报道 | 某REO（迷迭香精油） | 7.32% | [144] |
| 其他研究报道 | R. officinalis 1精油 | 13.3±2.9% | [145] |
| 其他研究报道 | R. officinalis 2精油 | 7.1±1.3% | [145] |
| 其他研究报道 | Ro样品精油 | 13.07% | [146] |
| 其他研究报道 | 某迷迭香精油 | 22.57% | [147] |
| 其他研究报道 | 某迷迭香精油 | 15% | [148] |
| 其他研究报道 | 部分样品 | 2–5% | [134] |
| 其他研究报道 | 部分样品 | 5.8–16.4% | [149] |
| 其他研究报道 | 迷迭香精油（范围） | 8.40–22.21% | [54] |
此外，提取方法中的微波辅助水蒸馏（MHDG）和亚临界流体萃取（SFME）可提高樟脑等含氧单萜化合物的含量，从而提升精油品质[106]；超临界萃取、水蒸气蒸馏（SD）和HYDRO法提取的精油中，樟脑均为主要成分[150]。在樟脑的提取与定量分析方面，可采用正戊烷提取迷迭香叶组织中的萜类化合物，以十三烷为内标，通过配备Elite-Betacydex β-环糊精毛细管柱的GC进行分析，利用保留时间与标准品比对鉴定樟脑，并通过内标法定量[65]。
超临界CO₂萃取中樟脑的提取参数与动力学特性
迷迭香精油中樟脑的提取方法、定量分析技术及超临界CO₂萃取参数与动力学特性研究显示，多种提取方法可影响樟脑的含量与提取效率，不同地理来源的迷迭香樟脑含量亦存在差异。定量分析方面，气相色谱-火焰离子化检测器（GC-FID）与气相色谱-质谱联用（GC-MS）是主要技术，通过与标准品（樟脑CAS号76-22-2）的保留时间或保留指数对比实现定性，采用外标法进行定量[151][111][112]。迷迭香精油的化学型分类中，樟脑型为已知的四种主要化学型之一。
不同提取方法及超临界CO₂萃取参数下的樟脑含量与提取效率对比
| 提取方法/参数条件 | 樟脑含量/提取效率 | 参考文献 |
|---|---|---|
| 水蒸气蒸馏（HD）（干基得率1.8%） | 樟脑含量达1.22% | [106] |
| 微波辅助水蒸气蒸馏（SFME，提取30分钟） | 樟脑含量为12.86% | [111] |
| 水蒸气蒸馏（HD，提取5分钟，峰值） | 樟脑含量达10.10% | [111] |
| 超临界CO₂萃取（S/F=1-3 g CO₂/g迷迭香） | 樟脑提取效率69-89% | [151] |
| 超临界CO₂萃取（床层表观密度0.48 g/cm³、孔隙率0.65，S/F=2.5 g CO₂/g迷迭香） | 樟脑含量约为10 wt.% | [151] |
| 超临界CO₂萃取（E-2床层，S/F<4） | 樟脑几乎耗尽 | [112] |
| 超临界CO₂萃取（E-1床层，S/F>4） | 樟脑耗尽（速率较慢） | [112] |
| 超临界CO₂萃取vs索氏提取法 | 前者樟脑与1,8-桉叶素总浓度约为后者18倍 | [112] |
| 西班牙型迷迭香 | 樟脑含量13-21% | [106] |
| 摩洛哥与突尼斯型迷迭香 | 樟脑含量5-15% | [106] |
研究表明，微波辅助水蒸气蒸馏（SFME）与HD所得精油的定性成分相似，但定量存在差异，HD提取樟脑含量在5分钟达峰值后随时间延长逐渐降低[111]；超临界CO₂萃取（SFE-CO₂）作为高效提取技术，其樟脑提取效率受溶剂与原料比（S/F）、床层几何结构影响显著[151][112]。
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